|Register||Search||Today's Posts||Mark Forums Read|
|DNA Extraction Forum DNA Isolation Forum. A forum specifically about questions on DNA extraction and purification.|
| ||LinkBack||Thread Tools||Display Modes|
DNAse in DNA Isolation Interfering with In Vitro Transcription Reaction
Hi, what i am doing is trying to remove all DNA template from my in vitro transcription reaction using T7 polymerase.
The DNAse is RNase free and DNAse digestion was at 37C but the problem is that in the first 20 minutes the DNA band intensity goes down when follow the DNA intensity on an agarose gel by time course!
Weirdly the DNA band intensity starts building up between 20minutes to 60minutes...
What is going on?
Re: DNAse in DNA Isolation Interfering with In Vitro Transcription Reaction
have you repeated this error? Can you show us an image of the gel?
The DNAse may be binding to the DNA and then interfering with gel migration and giving the appearance of reduced band intensity.
Possibly the DNAse was added at too high a concentration or the buffer in it affected the in vitro transcription.
I would try to order new DNase that is fresh DNAse from another company and try the experiment again.
Make sure you inactivate the DNAse first also.
|dna , dnase , interfering , isolation , reaction , transcription , vitro|
|Thread||Thread Starter||Forum||Replies||Last Post|
|Non-isotopic method for In vitro firstname.lastname@example.org||Protocols and Methods Forum||0||06-16-2008 02:15 PM|
|In vitro transcription problems||AudioHELP||Protocols and Methods Forum||2||12-17-2003 05:26 PM|
|T7 in vitro transcription||Vayuputra||Protocols and Methods Forum||0||07-19-2003 06:01 PM|